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InvisibleStipe-n CapMDiscord
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Re: Some notes about LCs [Re: Pastywhyte]
    #27029867 - 11/09/20 09:30 AM (3 years, 2 months ago)

I don't do a lot of LC but I have cooked lot's of agar. Never had a bad plate.

Edit* perhaps you are at a higher elevation out there in BC.


Edited by Stipe-n Cap (11/09/20 09:47 AM)


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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Stipe-n Cap] * 1
    #27029893 - 11/09/20 09:52 AM (3 years, 2 months ago)

I’m definitely at a higher elevation than most people.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Nurkurzda23]
    #27030452 - 11/09/20 03:59 PM (3 years, 2 months ago)

Quote:

Nurkurzda23 said:
I have read that a certain concentration of H2O2 in LC helps against contamination.  how much 3% solution do you use at approx 250 mL / 8.5 Oz?

are the jar lids are slightly open during they are autoclaved. (With tin foil over them) because I have a lot of contaminated jars... And don't know why




I've think read about 1% of the standard 3% solution from a long time  ago (used to be this guy who sold a H2O2 technique manual before it was common knowledge).  There seemed to be some disagreement when to add it.  I think the manual said before pressure cooking, but I read a recent thing on somewhere else (maybe this site) that was saying once the goes down to 140F.  I tried adding 1% at 140F and it was fine for transfers.  I didn't try to germinate on it, but I don't think you are supposed to use it for germinations.


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OfflineOm Namo Shivaya


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Re: Some notes about LCs [Re: Blue Helix]
    #27030460 - 11/09/20 04:07 PM (3 years, 2 months ago)

Hey all, i have not been able to find an answer on the forums for LC expansion and was wondering if any of you have experience with it.

I'm used to g2g and have recently swapped to LC. I've heard some say you should start every lc jar with a new wedge, because LC to LC can lead to senescence quickly.

Anyone have experience with expanding LC to LC and any experiences of senescence?

Thanks for any input


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Om Namo Shivaya] * 1
    #27030475 - 11/09/20 04:25 PM (3 years, 2 months ago)

Quote:

Om Namo Shivaya said:
Hey all, i have not been able to find an answer on the forums for LC expansion and was wondering if any of you have experience with it.

I'm used to g2g and have recently swapped to LC. I've heard some say you should start every lc jar with a new wedge, because LC to LC can lead to senescence quickly.

Anyone have experience with expanding LC to LC and any experiences of senescence?

Thanks for any input




I am sure someone here will inject and disagree, but I have not seen this rapid senescence that people warn about.  I usually refresh an LC every year, and it's done from a few ml of the last batch.  I'm not saying senescence isn't real--it is--but it didn't bother me at all.  Most of my grows are after one to three expansions.

If you are really worried about it, do what the agar people do (i.e. play endlessly with agar plates).  For me, though, the LC vacutainers are just too convenient, and I'm more into growing mushrooms than pretend playing microbiologist.


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OfflineGeinstein
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Re: Some notes about LCs [Re: Blue Helix]
    #27030838 - 11/09/20 09:50 PM (3 years, 2 months ago)

Quote:

Blue Helix said:
Quote:

Om Namo Shivaya said:
Hey all, i have not been able to find an answer on the forums for LC expansion and was wondering if any of you have experience with it.

I'm used to g2g and have recently swapped to LC. I've heard some say you should start every lc jar with a new wedge, because LC to LC can lead to senescence quickly.

Anyone have experience with expanding LC to LC and any experiences of senescence?

Thanks for any input




I am sure someone here will inject and disagree, but I have not seen this rapid senescence that people warn about.  I usually refresh an LC every year, and it's done from a few ml of the last batch.  I'm not saying senescence isn't real--it is--but it didn't bother me at all.  Most of my grows are after one to three expansions.

If you are really worried about it, do what the agar people do (i.e. play endlessly with agar plates).  For me, though, the LC vacutainers are just too convenient, and I'm more into growing mushrooms than pretend playing microbiologist.



It all matters what your growing


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Nothing breads nothing


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OfflineNurkurzda23
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Re: Some notes about LCs [Re: Pastywhyte]
    #27031175 - 11/10/20 07:45 AM (3 years, 2 months ago)

Hi

Is it possible that some - a little bit mycelium float on top of the jar of it is strong colonized?


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OfflineA.k.aM
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Re: Some notes about LCs [Re: Nurkurzda23]
    #27031177 - 11/10/20 07:47 AM (3 years, 2 months ago)

Yeah if you leave it long enough the whole top of the broth will be thick myc.


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LAGM2020


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OfflineNurkurzda23
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Re: Some notes about LCs [Re: A.k.a]
    #27031242 - 11/10/20 08:48 AM (3 years, 2 months ago)

Im very thankful
Otherwise I would have trashed it😄


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OfflineGeinstein
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Re: Some notes about LCs [Re: Geinstein]
    #27031258 - 11/10/20 09:00 AM (3 years, 2 months ago)

Quote:

Geinstein said:
I'm prepping some LC jars 10 x 200ml of T2 and T3 plates(cleaned up from dirty ass print)
My LC ends up looking like this :



(Bad light making for bad photos, but basically looks like Cristal clear water)


Now the batch I made 2 days ago(I inoculate 3 days after PCing) have all got this piece of grain clump, I'd like to know if it'll still be fine to use



My recipe is:
1g BRF
0.5g Dextrose
10ml second brew black Tea
200ml water

My only difference with this one is I mixed everything together before PCing were normally I'd just add everything and then add water without mixing and PC it. Could the mixing be the reason for the clump and can it still be used?



Could some one help?


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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Geinstein]
    #27031268 - 11/10/20 09:07 AM (3 years, 2 months ago)

None of those look crystal clear to me, they all look very turbid.


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OfflineGeinstein
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Re: Some notes about LCs [Re: Pastywhyte]
    #27031388 - 11/10/20 10:27 AM (3 years, 2 months ago)

I know it looks like that in the photo but it really does look like water

The bottom it the one with clamps, I'm gone redo them rather, my conserns are on way they clamped up.


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OfflineAsuraS
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Re: Some notes about LCs [Re: Pastywhyte]
    #27031396 - 11/10/20 10:33 AM (3 years, 2 months ago)

Quote:

Pastywhyte said:
None of those look crystal clear to me, they all look very turbid.




Maybe the BRF :shrug: Stuff works great but impedes visibility even
at very low amounts. It doesn't clear over time either...for me at least.


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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Asura]
    #27031401 - 11/10/20 10:37 AM (3 years, 2 months ago)

I love brf but the way I make LC traps the sediment, I just don’t trust turbid looking LC.


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InvisibleJosex
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Re: Some notes about LCs [Re: Asura]
    #27031403 - 11/10/20 10:38 AM (3 years, 2 months ago)

BRF LC will clear up too at a 0.2% ratio, IME. The uncolonized liquid will be like plain water when the LC is done. Flours are fun and myc loves to have some nice sediment to hold onto, but if I have LME that's what I'll use for sure.


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Asura]
    #27031558 - 11/10/20 12:28 PM (3 years, 2 months ago)

Quote:

Asura said:
Quote:

Pastywhyte said:
None of those look crystal clear to me, they all look very turbid.




Maybe the BRF :shrug: Stuff works great but impedes visibility even
at very low amounts. It doesn't clear over time either...for me at least.




You guys understand that when I say a "clear LC" I mean it settles when you stop stirring it, right?  No LCs are not crystal clear when being spun or swished around.  What I am talking about is the fluid between the blobs of mycelium (or slush if you are using a high-speed spinner) is clear.  I don't mean the whole LC is clear.  If you leave it alone for a few hours, the top should clear up.  If it doesn't, that usually means something is wrong.  It means that there are particles of something being suspended in there by the action of bacteria usually (I guess if the particles were SUPER small it could be Brownian motion too but I don't use stuff that fine in my LCs).


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Invisiblecoversall
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Re: Some notes about LCs [Re: Blue Helix]
    #27031591 - 11/10/20 12:52 PM (3 years, 2 months ago)

I PC'd two batches of LME broth the other day, and both formed sediment while in the PC. One batch was PC'd with the jars about the water and the other was done with the water at the same level as the water in the jars as I heard this can reduce sediment.

Is sediment normal/ok with LME broth? Will post pics when I get a chance to take some.


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OfflineAsuraS
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Re: Some notes about LCs [Re: coversall]
    #27031598 - 11/10/20 12:55 PM (3 years, 2 months ago)

One thing I think that helps with sediment is making sure the water in the PC
is above the water line in the jars. This reduces the amount of sediment
that gets caked on the side (which is usually none if you do this).


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OfflineBlue Helix
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Re: Some notes about LCs [Re: Asura]
    #27031602 - 11/10/20 12:59 PM (3 years, 2 months ago)

Quote:

Asura said:
One thing I think that helps with sediment is making sure the water in the PC
is above the water line in the jars. This reduces the amount of sediment
that gets caked on the side (which is usually none if you do this).




Wow, I never heard of this.  I had always allowed the water line in the PC to be below the bottom of the jars or very close to it.  With an AA and toggle switch (peacock) the cooker allows almost no steam to escape, so you can use very low water without worrying about it boiling dry.  I'll have to give the super high water a try with the new jars!  Thanks!


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OfflineBlue Helix
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Re: Some notes about LCs [Re: coversall]
    #27031609 - 11/10/20 01:03 PM (3 years, 2 months ago)

Quote:

coversall said:
I PC'd two batches of LME broth the other day, and both formed sediment while in the PC. One batch was PC'd with the jars about the water and the other was done with the water at the same level as the water in the jars as I heard this can reduce sediment.

Is sediment normal/ok with LME broth? Will post pics when I get a chance to take some.




No only is it "okay" some folks PREFER that there are solids in there so they can serve as nucleation sites for the mycelium's flocculation.  That's a fancy way of saying they like that to LC goes from cloudy to clear as it completes, and they use the clearness as an indicator the LC is done.  In any case, those solids have little to no effect on the LC's function.  Not having the solids is a cosmetic preference at best, and I don't think you should worry about it really.


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